Sorghum is an important cereal with diverse phenolic compounds that have potential health promoting benefits. The current study comparatively characterized the phenolic contents of two novel black-seeded sorghum lines (SC84 and PI570481) using different extraction systems (water, ethanol and their acidified counterparts) and evaluated their antioxidant and anti-inflammatory activities. Phenolic compositions were determined by spectrophotometric assays and HPLC analysis. Antioxidant activities were assessed by radical scavenging effects on nitric oxide (NO) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals, and the oxygen radical absorbance capacity (ORAC). Anti-inflammatory capacity was estimated by measuring levels of pro-inflammatory markers produced by lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Results showed that effects of solvent types and HCl on extraction efficiency differed among phenolic compounds and sorghum samples. Tannins were the most dominant polyphenols in the studied extracts (11.11–136.11 mg epicatechin equivalent/g sorghum). Sorghum extracts exerted more potent scavenging activity on DPPH than NO radicals. In LPS-activated RAW 264.7 cells, sorghum extracts dose-dependently inhibited the production of NO, interleukin-6 (IL-6), and intracellular reactive oxygen species (ROS), with ethanolic extracts showing greater anti-inflammatory activity. Positive correlations were noted between tannin content and DPPH radical scavenging activity, and anti-inflammatory capacity. These results suggest the potential role of tannin-rich sorghum extracts against inflammation and associated diseases.

Sorghum [Sorghum bicolor(L.) Moench] grain yield is severely affected by abiotic and biotic stresses during post-flowering stages, which has been aggravated by climate change. New parental lines having genes for various biotic and abiotic stress tolerances have the potential to mitigate this negative effect. Field studies were conducted under irrigated and dryland conditions with 128 exotic germplasm and 12 adapted lines to evaluate and identify potential sources for post-flowering drought tolerance and stalk and charcoal rot tolerances. The various physiological and disease related traits were recorded under irrigated and dryland conditions. Under dryland conditions, chlorophyll content (SPAD), grain yield and HI were decreased by 9, 44 and 16%, respectively, compared to irrigated conditions. Genotype RTx7000 and PI475432 had higher leaf temperature and grain yield, however, genotype PI570895 had lower leaf temperature and higher grain yield under dryland conditions. Increased grain yield and optimum leaf temperature was observed in PI510898, IS1212 and PI533946 compared to BTx642 (B35). However, IS14290, IS12945 and IS1219 had decreased grain yield and optimum leaf temperature under dryland conditions. Under irrigated conditions, stalk and charcoal rot disease severity was higher than under dryland conditions. Genotypes IS30562 and 1790E R had tolerance to both stalk rot and charcoal rot respectively and IS12706 was the most susceptible to both diseases. PI510898 showed combined tolerance to drought and Fusarium stalk rot under dryland conditions. The genotypes identified in this study are potential sources of drought and disease tolerance and will be used to develop better adaptable parental lines followed by high yielding hybrids.

Pearl millet is a staple food for people in arid and semi-arid regions of Africa and South Asia due to its high drought tolerance and nutritional qualities. A better understanding of the genomic diversity and population structure of pearl millet germplasm is needed to support germplasm conservation and genetic improvement of this crop. Here we characterized two pearl millet diversity panels, (i) a set of global accessions from Africa, Asia, and the America, and (ii) a collection of landraces from multiple agro-ecological zones in Senegal. We identified 83,875 single nucleotide polymorphisms (SNPs) in 500 pearl millet accessions, comprised of 252 global accessions and 248 Senegalese landraces, using genotyping by sequencing (GBS) of PstI-MspI reduced representation libraries. We used these SNPs to characterize genomic diversity and population structure among the accessions. The Senegalese landraces had the highest levels of genetic diversity (π), while accessions from southern Africa and Asia showed lower diversity levels. Principal component analyses and ancestry estimation indicated clear population structure between the Senegalese landraces and the global accessions, and among countries in the global accessions. In contrast, little population structure was observed across in the Senegalese landraces collections. We ordered SNPs on the pearl millet genetic map and observed much faster linkage disequilibrium (LD) decay in Senegalese landraces compared to global accessions. A comparison of pearl millet GBS linkage map with the foxtail millet (Setaria italica) and sorghum (Sorghum bicolor) genomes indicated extensive regions of synteny, as well as some large-scale rearrangements in the pearl millet lineage. We identified 83,875 SNPs as a genomic resource for pearl millet improvement. The high genetic diversity in Senegal relative to other regions of Africa and Asia supports a West African origin of this crop, followed by wide diffusion. The rapid LD decay and lack of confounding population structure along agro-ecological zones in Senegalese pearl millet will facilitate future association mapping studies. Comparative population genomics will provide insights into panicoid crop evolution and support improvement of these climate-resilient crops.

Dissecting the genetic architecture of stress tolerance in crops is critical to understand and improve adaptation. In temperate climates, early planting of chilling-tolerant varieties could provide longer growing seasons and drought escape, but chilling tolerance (75,000 data points from ∼16,000 plots) in multi-environment field trials in Kansas under natural chilling stress (sown 30-45 days early) and normal growing conditions. Joint linkage mapping with early-planted field phenotypes revealed an oligogenic architecture, with 5-10 chilling tolerance loci explaining 20-41% of variation. Surprisingly, several of the major chilling tolerance loci colocalize precisely with the classical grain tannin (Tan1 and Tan2) and dwarfing genes (Dw1 and Dw3) that were under strong directional selection in the US during the 20th century. These findings suggest that chilling sensitivity was inadvertently selected due to coinheritance with desired nontannin and dwarfing alleles. The characterization of genetic architecture with NAM reveals why past chilling tolerance breeding was stymied and provides a path for genomicsenabled breeding of chilling tolerance.

Stomatal density (SD) and stomatal complex area (SCA) are important traits that regulate gas exchange and abiotic stress response in plants. Despite sorghum (Sorghum bicolor) adaptation to arid conditions, the genetic potential of stomata-related traits remains unexplored due to challenges in available phenotyping methods. Hence, identifying loci that control stomatal traits is fundamental to designing strategies to breed sorghum with optimized stomatal regulation. We implemented both classical and deep-learning methods to characterize genetic diversity in 311 grain sorghum accessions for stomatal traits at two different field environments. Nearly 12,000 images collected from abaxial and adaxial leaf surfaces revealed substantial variation in stomatal traits. Our study demonstrated significant accuracy between manual and deep-learning methods in predicting SD and SCA. In sorghum, SD was 32-39% greater on the abaxial vs. the adaxial surface, while SCA on the abaxial surface was 2-5% lower than on the adaxial surface. GWAS identified 71 genetic loci (38 were environment-specific) with significant genotype to phenotype associations for stomatal traits. Putative causal genes underlying the phenotypic variation were identified. Accessions with similar SCA but carrying contrasting haplotypes for SD were tested for stomatal conductance and carbon assimilation under field conditions. Our findings provide a foundation for further studies on the genetic and molecular mechanisms controlling stomata patterning and regulation in sorghum. An integrated physiological, deep learning, and genomic approach allowed us to clarify the genetic control of natural variation in stomata traits in sorghum, and can be applied to other plants.

Sorghum is one of the most important food and feed cereal crops and has been gaining industrial importance in recent years for its biofuel, nutraceutical and antioxidant values. A genetic profile variation study was undertaken for the accumulation of phytochemicals in 61 diverse sorghum accessions differing in their growth habitat and grain color through non-targeted Gas Chromatography–Mass Spectrometry (GC-MS/MS) analysis. Mass Spectrometry–Data Independent AnaLysis (MS-DIAL) and MetaboAnalyst identified 221 metabolites belonging to 27 different phytochemicals. Tropical and temperate sorghums were distinct in their metabolic profiles with minimum overlaps, and 51 different metabolites were crucial in differentiating the two groups. Temperate sorghums had the ability to accumulate more of phenolic acids, phytosterols, flavonoids, carotenoids, and tropical sorghums for stress-related amino acids, sugars and fatty acids. Grain-color-based Partial Least Square–Discriminant Analysis (PLS-DA) analysis identified 94 Variable Importance in Projections (VIP) metabolites containing majority of flavonoids, phenylpropanoids and phytosterols. This study identified two sorghum lines (IS 7748 and IS 14861) with rich amounts of antioxidants (catechins and epicatechins) belonging to the group of condensed tannins that otherwise do not accumulate commonly in sorghum. Out of 13 metabolic pathways identified, flavonoid biosynthesis showed the highest expression. This study provided new opportunities for developing biofortified sorghum with enhanced nutraceutical and therapeutics through molecular breeding and metabolic engineering.